It is possible for your MyGo Pro to choose the optimum integration time needed to gain the maximum amount of data out of a reaction without saturating during a melt. Optimize Melt Acquisitions gives you the option to do this is under the Run Settings tab, please refer to Run Settings if you are unsure of the location of this setting.
Optimize Melt Acquisitions
To perform Optimize Melt Acquisitions you will need to perform a melt, please refer to Profile if you are unsure how to do this. This optimum value will be presented to you as an Automatic Integration Time found in the Experiment Log.
Once a reaction has completed you can find the automatic integration time by selecting Show log… at the bottom left of the screen as shown below.
Once selected you will be presented with the Experiment Log window as shown below. The automatic integration time for this specific experiment is 0.8s. This means that if this run was to be repeated then an integration time of over 0.8s would saturate the sensor.
Automatic Integration Time
This Automatic Integration Time, can be used in subsequent cycling runs. This means that if an experiment with the same reagents, concentrations and volumes is repeated, more optical data can be acquired throughout the run. The optimised integration time can be programmed into a new run under Run Settings, in Use Advanced Settings. These settings are useful when performing demanding applications, however the default Optical Settings are sufficient for most applications.
In the example below the Integration Time has been programed at 0.8s. This means that the instrument will acquire data for 0.8s per acquisition at every cycle.
Acquisitions per Cycle
This is the number of acquisitions the instrument will take at every cycle. The more acquisitions the instrument takes the more data points it has to average over for that cycle. The number of acquisitions depends on how long you want the reaction to be. Data quality will improve with increased number of acquisitions, however this may eventually become insignificant with very long runs. In the example below, 5 acquisitions will be taken per cycle. This means that every cycle will have a total integration time of 4s.
Optimise Melt Acquisitions in Advanced settings
If advanced optical settings have been programmed then the instrument will by default use these settings for cycling and melting. If Optimise Melt Acquisitions has been selected within advanced settings then the instrument will use the settings you have selected for integration time and acquisition per cycle for cycling; however will search for an optimum integration time for the melt.
Saturation and Example Settings
If you are using advanced settings and your run is saturating then you may want to decrease the integration time. Always refer to Show log… and see what the automatic integration time is.
Advanced settings are used mostly when performing multiplex reactions. With balanced probe concentrations and reactions containing red dyes, integration times of 1-2s is recommended. Integration times over 2s are not recommended as this will increase the chance of saturation and the likelihood of introducing undesirable optical artefacts.